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Application of AtNRT1.8 gene to strengthening resistance of crops to stress of heavy metals or salts

A technology of NRT1.8 and heavy metals, applied in the fields of plant bioengineering and plant improvement genetic engineering

Inactive Publication Date: 2012-07-04
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, there has not been any research or report involving the relationship between the AtNRT1.8 gene and the resistance of plants (especially crops) to heavy metals, salt or drought stress in the art

Method used

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  • Application of AtNRT1.8 gene to strengthening resistance of crops to stress of heavy metals or salts
  • Application of AtNRT1.8 gene to strengthening resistance of crops to stress of heavy metals or salts
  • Application of AtNRT1.8 gene to strengthening resistance of crops to stress of heavy metals or salts

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0114] Example 1. Regulatory effects of heavy metals and other stress factors on AtNRT1.8 gene

[0115] Through gene chip analysis, the inventors found that AtNRT1.8 was most strongly induced by cadmium among the 53 POT gene family members (data not shown). The sequence of the AtNRT1.8 gene used in this embodiment is as follows figure 1 shown.

[0116] In order to confirm whether this gene is induced by cadmium as shown in the gene chip, and further reveal whether it is induced by other stress factors, a Northern hybridization test (test method: Northern hybridization in classic plant molecular biology, refer to "Molecular Cloning Experiments") Guide, pp. 532-552).

[0117] The above test results show that the expression of AtNRT1.8 gene induced by cadmium has time- and concentration-dependent effects (see figure 2 A), and basically not regulated by the secondary effects of cadmium stress, osmotic stress and oxidative stress (such as figure 2 As shown in B, PEG is a hi...

Embodiment 2

[0119] Embodiment 2.AtNRT1.8 gene encodes introversion low-affinity nitrate transporter (Nitrate transporter)

[0120] The AtNRT1.8 gene belongs to the PTR gene family, also known as the NRT1 family or the POT family. Since this family of genes has been found to encode both nitrate transporter and oligopeptide transporter, in order to detect the transport substrate of the protein encoded by the AtNRT1.8 gene, the inventors studied AtNRT1 by injecting Xenopus oocytes. In vitro transport activity of 8 genes.

[0121] Test method: The coding sequence of AtNRT1.8 gene and the UTR sequences at both ends were subcloned into pGEMHE vector, transcribed into cRNA in vitro, injected into Xenopus oocytes (obtained from Capital Normal University), and electrophysiologically analyzed (voltage clamp Purchased from Dagan Company, the model is TEV-200A VOLTAGECLAMP). Test results such as image 3 shown.

[0122] The results showed that the protein encoded by AtNRT1.8 gene specifically...

Embodiment 3

[0123] Example 3. Expression and localization of AtNRT1.8 gene in Arabidopsis

[0124] Determining the tissue expression characteristics and subcellular localization of genes plays an important role in clarifying the mechanism of action of genes. Therefore, the inventors further studied the expression and localization of the AtNRT1.8 gene in Arabidopsis.

[0125] Referring to the method described in Dawar Hussain et al., The Plant Cell "Plant Cell", Volume 16, 1327-1339, the promoter region of the gene is fused with the reporter gene GUS, or the coding frame of the gene is fused with the reporter gene GFP for Transform Arabidopsis or onion epidermal cells. In situ hybridization, the forward sequence and reverse sequence of AtNRT1.8 were used to hybridize with the mRNA of tissue native. Test results such as Figure 4 shown.

[0126] The results showed: GUS histochemical analysis showed that the expression of AtNRT1.8 gene was mainly expressed in the vascular system of pla...

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Abstract

The invention relates to application of an AtNRT1.8 gene to strengthening the resistance of crops to the stress of heavy metals or salts, in particular to application of an AtNRT1.8 gene or encoded protein or polypeptide thereof to strengthening the resistance of plants to the stress of heavy metals, salts or draught. The invention also provides a genetic engineering method for strengthening the resistance of plants to the stress of heavy metals, salts or draught by utilizing the AtNRT1.8 gene, a vector and a host cell containing the AtNRT1.8 gene, a method for preparing transgenosis plants and the transgenosis plants. The method can strength the resistance of the plants to the stress of the heavy metals, the salts and the draught and has active effects for improving the yield and properties of crops and wide application potential.

Description

technical field [0001] The invention relates to the fields of plant bioengineering and plant improvement genetic engineering. Specifically, the present invention relates to the use of the AtNRT1.8 gene in enhancing the resistance of crops to heavy metals, salt or drought stress, and the method for improving the resistance of plants to heavy metals, salt or drought stress by transferring the gene and transgenic plants. Background technique [0002] The important reason why heavy metal cadmium pollution causes poisonous damage to plants is that it significantly inhibits the important process of nitrogen absorption and assimilation, resulting in nitrogen deficiency in plants and protein degradation in the body. Therefore, a direct consequence for crops is severe yield reduction. In addition, the accumulation of heavy metals in the edible parts of crops, such as rice, will also lead to a decrease in safety and quality, causing great safety hazards to people or animals who eat t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82A01H5/00C12N15/63C12N1/19C12N1/21C12N5/10
Inventor 龚继明
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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